Hi, all,
I am doing plaque lifts for a cDNA library screen. After the first set
of Hybond N membranes had been detected with DIG-labeled probes I had a
very low signal to background ratio.
When I asked a colleague for possible improvements he pointed out that I
took the wrong membrane (although there are certanly more reasons for
background) and that I should try a positively charged one. I then asked
three more coworkers and in the end it was two for Hybond N + and two
for Hybond N (uncharged). Amersham recommends Hybond N+ more than the
uncharged version and Boehringer told me to take uncharged membranes
rather than charged ones.
To make the confusion complete, I would like to ask the rest of you
which membrane to prefer.
Thanks Frank
P.S.
I am aware that the choice of membrane is not the only possibility for
background reduction and I try several other methods as well. But I am
just curious about the membrane issue.